DNA

Part:BBa_K3791019:Design

Designed by: Laura Sánchez Ruiz, Auba Fuster Palŕ   Group: iGEM21_UPF_Barcelona   (2021-10-12)


Efficient gRNA Spectinomycin


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

The gRNA architecture includes two DR separated by the spacer with 4 additional base pairs downstream. So the final structure is: repeat + spacer + 4 nucleotides + repeat.


Source

The repeat sequence was extracted from the parts registry (BBa_K2927006), whereas the spacer one was designed with the constraint that it had to be complementary to the sequence of the spectinomycin-resistance gene aiming to be detected (as explained in its own part page: BBa_K3791004).

References

[1] Campa, C. C., Weisbach, N. R., Santinha, A. J., Incarnato, D., & Platt, R. J. (2019). Multiplexed genome engineering by Cas12a and CRISPR arrays encoded on single transcripts. Nature Methods, 16(9), 887–893. https://doi.org/10.1038/s41592-019-0508-6
[2] Paul, B., & Montoya, G. (2020). CRISPR-Cas12a: Functional overview and applications. Biomedical Journal, 43(1), 8–17. https://doi.org/10.1016/j.bj.2019.10.005